Acute exercise-induced release of innate immune proteins via small extracellular vesicles changes with aerobic fitness and age.

AIM: Physical exercise triggers the secretion of small extracellular vesicles (sEVs) into the circulation in humans, enabling signalling crosstalk between tissues. Exercise-derived EVs and their cargo have been proposed to mediate adaptations to exercise; however, our understanding of how exercise-derived EV protein cargo is modulated by factors such as aerobic fitness and age of an individual is currently unknown. Here, we examined the circulating sEV proteome following aerobic exercise in healthy males of different ages and aerobic fitness to understand exercise-induced EV response during the aging process. METHODS: Twenty-eight healthy men completed a bout of 20-min cycling exercise at 70% estimated VO2peak . Small EVs were isolated from blood samples collected before and immediately after exercise, and then quantified using particle analysis and Western blotting. Small EV proteome was examined using quantitative proteomic analysis. RESULTS: We identified a significant increase in 13 proteins in small plasma EVs following moderate-to-vigorous intensity exercise. We observed distinct changes in sEV proteome after exercise in young, mature, unfit, and fit individuals, highlighting the impact of aerobic fitness and age on sEV protein secretion. Functional enrichment and pathway analysis identified that the majority of the significantly altered sEV proteins are associated with the innate immune system, including proteins known to be damage-associated molecular patterns (DAMPs). CONCLUSION: Together, our findings suggest that exercise-evoked acute stress can positively challenge the innate immune system through the release of signalling molecules such as DAMPs in sEVs, proposing a novel EV-based mechanism for moderate-to-vigorous intensity exercise in immune surveillance pathways.

Exercise increases the release of NAMPT in extracellular vesicles and alters NAD+ activity in recipient cells.

Aging is associated with a loss of metabolic homeostasis, with cofactors such as nicotinamide adenine dinucleotide (NAD+ ) declining over time. The decrease in NAD+ production has been linked to the age-related loss of circulating extracellular nicotinamide phosphoribosyltransferase (eNAMPT), the rate-limiting enzyme in the NAD+ biosynthetic pathway. eNAMPT is found almost exclusively in extracellular vesicles (EVs), providing a mechanism for the distribution of the enzyme in different tissues. Currently, the physiological cause for the release of eNAMPT is unknown, and how it may be affected by age and physical exercise. Here, we show that release of small EVs into the bloodstream is stimulated following moderate intensity exercise in humans. Exercise also increased the eNAMPT content in EVs, most prominently in young individuals with higher aerobic fitness. Both mature fit and young unfit individuals exhibited a limited increase in EV-eNAMPT release following exercise, indicating that this mechanism is related to both the age and physical fitness of a person. Notably, unfit mature individuals were unable to increase the release of eNAMPT in EVs after exercise, suggesting that lower fitness levels and aging attenuate this important signalling mechanism in the body. EVs isolated from exercising humans containing eNAMPT were able to alter the abundance of NAD+ and SIRT1 activity in recipient cells compared to pre-exercise EVs, indicating a pathway for inter-tissue signalling promoted through exercise. Our results suggest a mechanism to limit age-related NAD+ decline, through the systemic delivery of eNAMPT via EVs released during exercise.

Effect of Acute Heart Rate Variability Biofeedback on H-reflex Modulation: A Pilot Study.

Heart rate variability biofeedback (HRV BFB) is paced breathing scheme that stimulates resonance in the cardiovascular system. This study aimed to investigate the effect of a single-session HRV BFB on Hoffman reflex (H-reflex) of the soleus muscle. Twelve healthy males (height: 173.7 ± 7.18 cm; weight: 72.7 ± 17.7 kg; age: 24.0 ± 5.02 yrs) completed a randomized-crossover intervention involving a 10-minute HRV BFB and normal breathing (CON) separated by 48 hours. Results revealed significantly lower 1a afferent activation after HRV BFB. Similarly, the HRV BFB also demonstrated lower proportion of activated motor neurons from 1a afferents. In conclusion, an acute HRV BFB influenced the reduction in motoneuron excitability at resting condition.

Effects of wrist position on eccentric exercise-induced muscle damage of the elbow flexors.

We tested the hypothesis that the magnitude of changes in indirect muscle damage markers would be greater after maximal elbow flexor eccentric exercise in the supinated (shorter biceps brachii) than neutral wrist (longer) position, and the difference in the magnitude would be associated with greater elongation over contractions for the supinated than neutral position, rather than the initial muscle length. Ten untrained men (21-39 years) performed two bouts of 10 sets of 6 maximal isokinetic eccentric contractions of the elbow flexors in the supinated position for one arm and neutral position for the other arm separated by 2 weeks in a randomized order. Biceps brachii myotendinous junction (MTJ) movements during eccentric contractions were recorded by B-mode ultrasonography, and the displacement from the start to end of each contraction was quantified. Peak torque (supinated: 367.8 ± 112.5 Nm, neutral: 381.5 ± 120.4 Nm) and total work (1816 ± 539 J, 1865 ± 673 J) produced during eccentric contractions were similar between conditions. The average MTJ displacement increased (P < .05) from the 1st set (8.0 ± 2.0 mm) to 10th set (15.8 ± 1.9 mm) for the supinated condition, but no such increase was found in the neutral condition (1st set: 5.1 ± 1.0 mm, 10th set: 5.0 ± 0.8 mm). Changes in indirect muscle damage markers (maximal voluntary isometric contraction torque, range of motion, serum creatine kinase activity, and muscle soreness) after exercise were greater (P < .05) for the supinated than neutral condition. These results suggest that the greater muscle damage marker changes for the supinated than neutral wrist position was associated with the greater muscle lengthening (strain).

Coronavirus (COVID-19), Coagulation, and Exercise: Interactions That May Influence Health Outcomes.

The proinflammatory cytokine storm associated with coronavirus disease 2019 (COVID-19) negatively affects the hematological system, leading to coagulation activation and endothelial dysfunction and thereby increasing the risk of venous and arterial thrombosis. Coagulopathy has been reported as associated with mortality in people with COVID-19 and is partially reflected by enhanced D-dimer levels. Poor vascular health, which is associated with the cardiometabolic health conditions frequently reported in people with severer forms of COVID-19, might exacerbate the risk of coagulopathy and mortality. Sedentary lifestyles might also contribute to the development of coagulopathy, and physical activity participation has been inherently lowered due to at-home regulations established to slow the spread of this highly infectious disease. It is possible that COVID-19, coagulation, and reduced physical activity may contribute to generate a "perfect storm," where each fuels the other and potentially increases mortality risk. Several pharmaceutical agents are being explored to treat COVID-19, but potential negative consequences are associated with their use. Exercise is known to mitigate many of the identified side effects from the pharmaceutical agents being trialled but has not yet been considered as part of management for COVID-19. From the limited available evidence in people with cardiometabolic health conditions, low- to moderate-intensity exercise might have the potential to positively influence biochemical markers of coagulopathy, whereas high-intensity exercise is likely to increase thrombotic risk. Therefore, low- to moderate-intensity exercise could be an adjuvant therapy for people with mild-to-moderate COVID-19 and reduce the risk of developing severe symptoms of illness that are associated with enhanced mortality.

Can Heart Rate Variability Biofeedback Improve Athletic Performance? A Systematic Review.

This systematic review was conducted to evaluate the effect of heart rate variability biofeedback (HRV BFB) on performance of athletes. Six electronic databases (Springerlink, SportDiscus, Web of Science, PROQUEST Academic Research Library, Google Scholar, and ScienceDirect) and article references were searched. Eligibility criteria were: 1. experimental studies involving athletes randomly allocated among groups (randomized control trial); 2. availability of HRV BFB as a treatment compared to a control condition (CON) that involves regular sport/dance training, a placebo (PLA) or other methods of BFB; 3. performance-related variables such as a dependent index; and, 4. peer-reviewed articles written in English. Out of 660 articles, six studies were included in the systematic review which involved 187 athletes (females: n = 89; males n = 98). Six studies compared HRV BFB with a CON, three studies compared HRV BFB with a PLA, and two studies differentiated HRV BFB with other methods of BFB. Findings support HRV BFB as a potential intervention to improve fine and gross motor function in athletes.

Acquired and Genetic Thrombotic Risk Factors in the Athlete.

While athletes are often considered the epitome of health due to their physique and lowered potential for metabolic and cardiovascular diseases, they may also be at risk for the onset and development of venous thromboembolism (VTE). In an attempt to achieve and remain competitive, athletes are frequently exposed to numerous athlete-specific risk factors, which may predispose them to VTE through the disruption of factors associated with Virchow's triad (i.e., hypercoagulability, venous stasis, and vessel wall injury). Indeed, hypercoagulability within an athletic population has been well documented to occur due to a combination of multiple factors including exercise, dehydration, and polycythemia. Furthermore, venous stasis within an athletic population may occur as a direct result of prolonged periods of immobilization experienced when undertaking long-distance travels for training and competition, recovery from injury, and overdevelopment of musculature. While all components of Virchow's triad are disrupted, injury to the vessel wall has emerged as the most important factor contributing to thrombosis formation within an athletic population, due to its ability to influence multiple hemostatic mechanisms. Vessel wall injury within an athletic population is often related to repetitive microtrauma to the venous and arterial walls as a direct result of sport-dependent trauma, in addition to high metabolic rates and repetitive blood monitoring. Although disturbances to Virchow's triad may not be detrimental to most individuals, approximately 1 in 1,000 athletes will experience a potentially fatal post-exercise thrombotic incidence. When acquired factors are considered in conjunction with genetic predispositions to hypercoagulability present in some athletes, an overall increased risk for VTE is present.

Time of day and short-duration high-intensity exercise influences on coagulation and fibrinolysis.

Exercise has been demonstrated to have considerable effects upon haemostasis, with activation dependent upon the duration and intensity of the exercise bout. In addition, markers of coagulation and fibrinolysis have been shown to possess circadian rhythms, peaking within the morning (0600-1200 h). Therefore, the time of day in which exercise is performed may influence the activation of the coagulation and fibrinolytic systems. This study aimed to examine coagulation and fibrinolytic responses to short-duration high-intensity exercise when completed at different times of the day. Fifteen male cyclists (VO2max: 60.3 ± 8.1 ml kg-1 min-1) completed a 4-km cycling time trial (TT) on five separate occasions at 0830, 1130, 1430, 1730 and 2030. Venous blood samples were obtained pre- and immediately post-exercise, and analysed for tissue factor (TF), tissue factor pathway inhibitor (TFPI), thrombin-anti-thrombin complexes (TAT) and D-Dimer. Exercise significantly increased plasma concentrations of TF (p < .0005), TFPI (p < .0006), TAT complexes (p < .0012) and D-Dimer (p < .0003). There was a time-of-day effect in pre-exercise TF (p = .004) and TFPI (p = .031), with 0830 greater than 1730 (p .001), while 1730 was less than 2030 h (p = .008), respectively. There was no significant effect of time of day for TAT (p = .364) and D-Dimer (p = .228). Power output, TT time and heart rate were not significantly different between TTs (p > .05); however, percentage VO2max was greater at 1730 when compared to 2030 (p = .04). Due to a time-of-day effect present within TF, peaking at 0830, caution should be applied when prescribing short-duration high-intensity exercise bout within the morning in populations predisposed to hypercoagulability.

Assessment of Muscle Pain Induced by Elbow-Flexor Eccentric Exercise.

CONTEXT: Delayed-onset muscle soreness (DOMS) is a common muscle pain that many people experience and is often used as a model of acute muscle pain. Researchers have reported the effects of various interventions on DOMS, but different DOMS assessment protocols used in these studies make it difficult to compare the effects. OBJECTIVE: To investigate DOMS characteristics after elbow-flexor eccentric exercise to establish a standardized DOMS assessment protocol. DESIGN: Descriptive laboratory study. SETTING: Research laboratory. PATIENTS OR OTHER PARTICIPANTS: Ten healthy, untrained men (21-39 years). INTERVENTION(S): Participants performed 10 sets of 6 maximal isokinetic eccentric contractions of the elbow flexors. MAIN OUTCOME MEASURE(S): Indirect muscle-damage markers were maximal voluntary isometric contraction torque, range of motion, and serum creatine kinase activity. Muscle pain was assessed before exercise, immediately postexercise, and 1 to 5 days postexercise using (1) a visual analog scale (VAS), (2) a category ratio-10 scale (CR-10) when applying static pressure and palpation at different sites (3, 9, and 15 cm above the elbow crease), and (3) pressure-pain thresholds (PPTs) at 50 sites (pain mapping). RESULTS: Maximal voluntary isometric contraction and range of motion decreased and creatine kinase activity increased postexercise, indicating muscle damage. Palpation induced greater pain than static pressure, and longitudinal and transverse palpations induced greater pain than circular palpation (P < .05). The PPT was lower in the medial region before exercise, but the pain-sensitive regions shifted to the central and distal regions of the biceps brachii at 1 to 3 days postexercise (P < .05). The VAS was correlated with the CR-10 scale (r = 0.91, P < .05) but not with the PPT (r = -0.28, P = .45). CONCLUSIONS: The way in which muscles are assessed affects the pain level score. This finding suggests that pain level and pain threshold cannot be used interchangeably and that the central and distal regions of the biceps brachii should be included in DOMS assessment using the VAS, CR-10 scale, and PPT after elbow-flexor eccentric exercise.

Reduced muscle lengthening during eccentric contractions as a mechanism underpinning the repeated-bout effect.

This study investigated biceps brachii distal myotendinous junction (MTJ) displacement during maximal eccentric elbow flexor contractions to test the hypothesis that muscle length change would be smaller (less MTJ displacement) during the second than the first exercise bout. Ten untrained men performed two eccentric exercise bouts (ECC1 and ECC2) with the same arm consisting of 10 sets of six maximal isokinetic (60°/s) eccentric elbow flexor contractions separated by 4 wk. Biceps brachii distal MTJ displacement was assessed using B-mode ultrasonography, and changes in the displacement (muscle length change) from the start to the end of each contraction during each set and over 10 sets were compared between bouts by two-way repeated-measures ANOVA. Several indirect muscle damage markers were also measured and compared between bouts by two-way repeated-measures ANOVA. The magnitude of MTJ displacement (average of six contractions) increased from set 1 (8.2 ± 4.7 mm) to set 10 (16.4 ± 4.7 mm) during ECC1 (P < 0.05), but no significant changes over sets were evident during ECC2 (set 1: 8.5 ± 4.0 mm; set 10: 9.3 ± 3.1 mm). Changes in maximal voluntary isometric contraction strength, range of motion, muscle thickness, ultrasound echo intensity, serum creatine kinase activity, and muscle soreness (visual analog scale) were smaller (P < 0.05) following ECC2 than ECC1, showing less damage in the repeated bout. These results indicate that the magnitude of muscle lengthening was less during the second than the first eccentric exercise bout, which appears to be a mechanism underpinning the repeated-bout effect.

Pacing strategies during the swim, cycle and run disciplines of sprint, Olympic and half-Ironman triathlons.

PURPOSE: This study investigated the influence of distance on self-selected pacing during the swim, cycle and run disciplines of sprint, Olympic and half-Ironman (HIM) distance triathlon races. METHOD: Eight trained male triathletes performed the three individual races in <2 months. Participants' bikes were fitted with Schoberer Rad Meßtechnik to monitor speed, power output and heart rate during the cycle discipline. Global positioning system was worn to determine speed and heart rate during the swim and run disciplines. RESULT: An even swim pacing strategy was adopted across all distances. A more stochastic pacing was observed during the HIM cycle [standard deviation of exposure variation analysis (EVASD) = 3.21 ± 0.61] when compared with the sprint cycle discipline (EVASD = 3.84 ± 0.44, p = 0.018). Only 20.9 ± 4.1 % of the cycling time was spent more than 10 % above the mean power output in the HIM, compared with 43.8 ± 2.9 % (p = 0.002) and 37.7 ± 11.1 % (p = 0.039) during the sprint and Olympic distance triathlons, respectively. Conversely, 13.6 ± 5.1 % of the cycling time was spent 5-10 % below the mean power output during the HIM, compared with 5.9 ± 1.2 % (p = 0.034) and 8.0 ± 5.1 % (p = 0.045) during the sprint and Olympic distance triathlons, respectively. A negative pacing strategy was adopted during the sprint distance run, compared with positive pacing strategy during the Olympic and HIM. CONCLUSION: Results of this study suggest that pacing strategies during triathlon are highly influenced by distance and discipline, and highlight the importance of developing pacing strategies based on distance, strengths and individual fitness.

Changes in electrical pain threshold of fascia and muscle after initial and secondary bouts of elbow flexor eccentric exercise.

PURPOSE: This study investigated changes in electrical pain threshold (EPT) after repeated eccentric exercise bouts to test the hypothesis that fascia would become more sensitive than muscle when greater delayed onset muscle soreness (DOMS) is induced. METHODS: Ten young men performed two eccentric exercise bouts (ECC1, ECC2) consisting of ten sets of six maximal isokinetic eccentric contractions of the elbow flexors with the same arm separated by 4 weeks. Maximal voluntary isometric contraction torque, range of motion, muscle soreness assessed by a visual analogue scale (VAS) and pressure pain threshold (PPT) were measured before, immediately after and 1-5 days after exercise. EPT was assessed in the biceps brachii fascia (BBF), biceps brachii muscle, and brachialis fascia (BF) 1 day before, immediately after, and 1, 2 and 4 days after exercise. RESULTS: All measures showed smaller changes (P < 0.05) after ECC2 than ECC1. EPT decreased after both bouts and the largest decreases were evident at 2 days post-exercise (P < 0.05). The decreases in EPT after ECC1 were greater (P < 0.05) for both BBF (Baseline: 1.45 ± 0.23 mA, 2 days post-exercise: 0.13 ± 0.11 mA) and BF (1.64 ± 0.29 mA, 0.26 ± 0.2 mA) than muscle (1.56 ± 0.29 mA, 0.69 ± 0.33 mA). Changes in EPT were correlated with the changes in PPT (r = 0.63-0.87, P ≤ 0.05) but not with VAS (r = -0.01 to 0.50). CONCLUSION: These results show that fascia becomes more sensitive than muscle to electrical stimulation after the initial eccentric exercise, suggesting that damage inflammation to fascia than muscle fibres is more associated with DOMS.